US10017573B2 - Anti-αβTCR antibody - Google Patents

Anti-αβTCR antibody Download PDF

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US10017573B2
US10017573B2 US14/241,099 US201214241099A US10017573B2 US 10017573 B2 US10017573 B2 US 10017573B2 US 201214241099 A US201214241099 A US 201214241099A US 10017573 B2 US10017573 B2 US 10017573B2
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antibody
antibodies
cells
bma031
cell
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Daniel Snell
Andreas Menrad
Gina Lacorcia
Srinivas Shankara
Huawei Qiu
Clark Pan
Benjamin Kebble
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Genzyme Corp
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2809—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against the T-cell receptor (TcR)-CD3 complex
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2893—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD52
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1138—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against receptors or cell surface proteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/71—Decreased effector function due to an Fc-modification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • the present invention relates to an antibody specific for the alpha beta T cell receptor ( ⁇ TCR).
  • ⁇ TCR alpha beta T cell receptor
  • the invention relates to a humanized anti- ⁇ TCR antibody, which is derived from the murine monoclonal antibody BMA031, and the use of said humanized antibody in immunosuppressive therapy.
  • polyclonal antisera against lymphocytes is well known for the purpose of immunosuppression.
  • antisera are labor-intensive to produce, show properties which vary between batches, and the specificity which can be obtained using polyclonal antisera is limited.
  • OKT3 has been extensively used for the treatment of patients undergoing acute allograft rejection after kidney transplantation (Russell, P. S., Colvin, R. B., Cosimi, A. B. (1984) Annu. Rev. Med. 35:63 and Cosimi, A. B., Burton, R. C., Colvin, R. B. et al. (1981) Transplantation 32:535).
  • OKT3 and rabbit complement were used for purging mature T cells from donor marrow to prevent acute graft versus host disease (GVHD) in allogeneic bone marrow transplantation (Prentice, H. G., Blacklock, H. A., Janossy, G. at al.
  • the T3-antigen complex recognized by CD3-mAbs (e.g., OKT3) is postulated to play a crucial role during T cell activation.
  • Alpha/beta T lymphocytes recognize peptide—MHC ligands by means of a multimeric protein ensemble termed the ⁇ T cell antigen receptor (TCR).CD3 complex.
  • TCR ⁇ T cell antigen receptor
  • This structure is composed of a variable ⁇ TCR dimer which binds antigens and three invariant dimers (CD3 ⁇ , ⁇ and ⁇ ) which are involved in TCR.CD3 surface transport, stabilization and signal transduction.
  • the alpha beta T cell receptor ( ⁇ TCR) is expressed on the majority (approx. 95%) of T cells and has a critical role in T cell activation via engagement of antigen displayed on MHC.
  • ⁇ TCR gamma delta T cell receptor
  • the ⁇ TCR positive cell population plays an important role in the innate immune response in defense against opportunistic infections of bacterial, viral and fungal origin.
  • Gamma delta T cells do not play a role in graft rejection in transplantation. Therefore, targeting the ⁇ TCR positive cell population and sparing the ⁇ TCR positive population should allow for significant therapeutic efficacy whilst maintaining a baseline immune protection against opportunistic infections.
  • the mouse IgG2b monoclonal antibody BMA031 (Borst et al. (November 1990) Hum. Immunol. 29(3):175-88; EP0403156) is specific for the common determinant on the TCR alpha/beta/CD3 complex, and does not bind to the gamma-delta TCR.
  • BMA031 is highly immunosuppressive and is capable of inducing apoptosis of activated T cells via a mechanism of activation-induced cell death (AICD) (Wesselborg et al. (May 1993) J. Immunol. 150(10): 4338-4345).
  • AICD activation-induced cell death
  • BMA031 does not engage human Fc gamma receptors (Fc ⁇ R) in the majority of the human population (approximately 10% of human possess Fc ⁇ Rs which do bind to mouse IgG2b isotype). As such the antibody does not cause T cell activation via cross-linking of the T cell receptor and, therefore, it does not induce T cell activation or the associated cytokine release.
  • Fc ⁇ R human Fc gamma receptors
  • BMA031 is a murine antibody and, as such, is not suitable for repeat dosing in human subjects in view of the human anti-mouse antibody (HAMA) response elicited therein.
  • HAMA human anti-mouse antibody
  • EuCIV3 was originally generated on a wild-type human IgG1 or IgG4 backbone which still retains Fc ⁇ R binding. These humanized antibodies therefore allowed for T cell activation, proliferation and the concomitant cytokine release and as such were significantly different to the original properties of BMA031.
  • a humanized monoclonal antibody which comprises the CDRs of BMA031, and retains the binding affinity of BMA031 for its cognate antigen.
  • said humanized antibody comprises a heavy chain variable region comprising the CDRs set forth in SEQ ID NOs: 7, 12 or 13 and the human IGH3-23 framework set forth in SEQ ID NO: 17, wherein framework position 6 is a donor residue; in an alternative embodiment, framework position 18 is a donor residue.
  • framework positions 49 and/or 69 are donor residues.
  • the humanized monoclonal antibody comprises a heavy chain variable region comprising the CDRs set forth in SEQ ID NOs: 15 or 16 and the human IGHV1-3*01 framework set forth in SEQ ID NO: 18, wherein one or more of framework positions 38, 44 and/or 48 is a donor residue; in an alternative embodiment, framework positions 44 and 48 are donor residues.
  • the humanized monoclonal antibody comprises a light chain variable region comprising the CDRs set forth in SEQ ID NO: 14 and the human IGKV3-11*01 framework set forth in SEQ ID NO: 19, wherein framework positions 70 and/or 71 are donor residues.
  • position 46 is a donor residue.
  • antibodies according to the first embodiment include antibodies which comprise a heavy chain variable region selected from the heavy chains comprising the sequences set forth in SEQ ID NO: 7, SEQ ID NO: 12 and SEQ ID NO: 13, and a light chain variable region sequence comprising the sequence as set forth in SEQ ID NO: 14.
  • antibodies according to the second embodiment include antibodies which comprise a heavy chain variable region selected from the heavy chains comprising the sequences set forth in SEQ ID NO: 15 and SEQ ID NO: 16, and a light chain variable region comprising the sequence as set forth in SEQ ID NO: 14.
  • the humanized antibodies according to the described embodiments are humanized versions of the BMA031. Their primary structures differ from that of the humanized antibody EuCIV3, which has decreased binding to the ⁇ TCR as compared to BMA031.
  • CDRs are indicated by means of annotation or underlining.
  • Frameworks are all sequences outside of the CDRs, which are defined according to the “Kabat” numbering system and extended, where applicable, by use of “IMGT” CDR definition. If a framework residue is not indicated to be changed to match a donor sequence, it will ordinarily be understood to be an acceptor residue.
  • the humanized antibodies may comprise a constant region.
  • the constant region is of human origin.
  • the humanized antibodies of the invention may be further modified by Fc engineering.
  • Immunoglobulins are liable to cross-link Fc ⁇ receptors, which can lead to constitutive T cell activation for anti-T cell antibodies.
  • antibodies can be modified to remove the Fc region, such as by the generation of Fab or Fv fragments; however, truncated immunoglobulins lack beneficial effector functions and exhibit a lower serum half-life. Therefore, the Fc region of the humanized antibody can be modified to prevent Fc ⁇ cross-linking.
  • Exemplary techniques include generation of aglycosylated immunoglobulins, for instance by modification of the Fc region by an N297Q mutation.
  • Immunoglobulins which fail to bind Fc ⁇ are also described by Armour et al., (1999) Eur. J. Immunol. 29:2613-2624.
  • the modification effected to IgG1 is known as the ⁇ ab modification, and consists in a combination of the ⁇ a mutation, in which IgG residues are substituted at positions 327, 330 and 331, and IgG2 residues substituted at positions 233-236, and the ⁇ b mutation, in which residue 236 is deleted.
  • the glycosylation pattern of antibodies according to the invention can be modified.
  • the antibody comprised one or more of mutations S298N, T299A and Y300S.
  • the antibody comprises two or more of mutations N297Q, S298N, T299A and Y300S.
  • a humanized antibody comprising the multiple mutations N297Q/S298N/Y300S, S298N/T299A/Y300S or S298N/Y300S.
  • a humanized monoclonal antibody which comprises the CDRs of BMA031, and retains the T cell suppression properties of BMA031.
  • Said humanized antibody preferably comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NOs: 12, 13, 15 or 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 14.
  • nucleic acid encoding at least a heavy chain variable region of a humanized monoclonal antibody according to the preceding aspects of the described embodiments.
  • the nucleic acid may encode variable and constant regions of the humanized antibody.
  • Heavy and light chains may be encoded on separate nucleic acids or on the same nucleic acid molecule.
  • a cell which expresses a nucleic acid according to the preceding aspect.
  • the cell is, for example, a cell adapted to express antibody molecules in culture.
  • the nucleic acid may include signal sequences and/or other sequences or modifications which are required for, or which modulate, expression of the antibody molecule in the cell, and/or secretion of the antibody molecule from the cell.
  • a humanized antibody is provided as described in the foregoing aspects, for use in suppressing a T cell mediated response in a subject.
  • the T cell mediated response can be involved in a condition selected from tissue transplantation, including solid organ transplant and composite tissue transplant, tissue grafting, multiple sclerosis and type 1 diabetes.
  • another embodiment provides a method for treating a subject suffering from a condition involving an aberrant T cell mediated response comprising administering to a subject in need thereof a pharmaceutically effective dose of an antibody according to the described embodiments.
  • Humanized non-activatory anti- ⁇ TCR antibodies which do not induce cytokine release have thus been generated which are capable of selective modulation of the ⁇ TCR and of inducing apoptosis of activated ⁇ TCR positive T cells. These antibodies have been generated for use as immunosuppressive agents in T cell mediated diseases. These antibodies have been generated through humanization of a mouse anti- ⁇ TCR antibody BMA031 and by Fc-engineering of the humanized antibodies to prevent engagement of Fc gamma receptors. The antibodies according to the described embodiments retain the binding affinity of BMA031, unlike the humanized versions of BMA031 available in the art.
  • the immunosuppressive properties of antibodies according to the described embodiments are superior to those of BMA031. Moreover, unlike the humanized BMA031 antibodies of the prior art, the antibodies according to the described embodiments do not induce cytokine release in normal PBMC.
  • an antibody comprising a modified Fc, in which said modified Fc comprises a modified glycosylation pattern which reduces Fc ⁇ R receptor binding, comprising one or more of mutations S298N, T299A and Y300S.
  • the antibody comprises two or more of mutations N297Q, S298N, T299A and Y300S.
  • the antibody comprises the multiple mutations N297Q/S298N/Y300S, S298N/T299A/Y300S or S298N/Y300S.
  • the antibody may be an antibody as described in preceding aspects of the invention.
  • a multispecific antibody comprising at least a heavy chain of a first binding domain as described in the preceding aspects of the invention, and a second binding domain specific for a tumor-specific antigen.
  • the first binding domain comprises a heavy chain according to the second aspect of the invention.
  • the multispecific antibody can comprise many different conformations; in one embodiment, it comprises an anti-TCR/CD3 scFv and an anti-tumor scFv.
  • the multispecific antibody is bispecific.
  • FIG. 1 BMA031 binds more strongly to ⁇ TCR compared to EuCIV3.
  • EuCIV3 has a decreased potency compared to BMA031.
  • FIG. 2 EuCIV3 is less potent than BMA031 in an in vitro education (IVE) assay.
  • CD8+ T cells were treated with anti- ⁇ TCR antibodies at various concentrations (x-axis) and co-cultured with autologous dendritic cells pulsed with the CMV peptide 495-503 (pp65) for seven days.
  • FIG. 3 HEBE1 binds ⁇ TCR comparably to BMA031 in a competition assay.
  • EuCIV3 has a decreased potency compared to BMA031 and HEBE1.
  • FIG. 4 HEBE1 has similar potency to EuCIV3 in an in vitro education (IVE) assay.
  • the IVE assay was performed as described in respect of FIG. 2 .
  • FIG. 5 Schematic showing iterative rounds of mutagenesis of anti- ⁇ TCR variable domains.
  • Framework in shaded box depicts the FR region where certain mouse residues lie. Shaded residues in first row of mutations are the mouse amino acids that are useful to maintain off-rate. Shaded residues in second row of mutations are the mouse residues surrounding the CDR regions which were retained during the final germlining process.
  • FIG. 6 Optimized humanized antibody has improved off-rate compared to BMA031.
  • BMA031 had a better off-rate compared to EuCIV3 and HEBE1.
  • By optimizing the binding domain of HEBE1 we were able to improve the off-rate of HEBE1.H10 compared to BMA031.
  • FIG. 7 Optimized humanized antibody has improved off-rate compared to BMA031.
  • FIG. 8 Optimized humanized antibody has improved off-rate compared to BMA031 in both ⁇ ab and aglycosylated formats.
  • the kinetics of antibody dissociation from ⁇ TCR on T cells was measured by flow cytometry.
  • FIG. 9 Optimization of HEBE1 leads to equivalent functionality as BMA031.
  • BMA031 inhibited education of CD8+ T cells, as they were unable to lyse specific targets in a dose dependent manner.
  • the parental humanized antibody, HEBE1 was not as potent as BMA031 and was able to only inhibit education at the highest dose (similar results observed with a second non-improved humanized Ab, HEBE1 H13). Further improvements were made to the humanized antibody, HEBE1 H10, which had equivalent potency to BMA031 in this assay.
  • FIG. 10 IVE data with anti- ⁇ TCR antibodies.
  • FIG. 11 Antigen positive cells from IVE assay as determined by antigen-specific tetramer binding.
  • IVE assay Cells which are antigen-positive (i.e., have been educated within IVE assay) are able to bind to an MHC-tetramer molecule.
  • IVE assay was conducted in the presence of antibody which has been able to prevent the education of T cells to antigen, there were fewer cells able to bind to the MHC-tetramer at the end of the assay.
  • FIG. 12 Proliferation of PBMCs in presence of anti- ⁇ TCR antibodies, OKT3 and stimulatory beads.
  • FIG. 13 Cytokine release from PBMCs in presence of anti- ⁇ TCR antibodies.
  • Cytokine release profile of anti- ⁇ TCR antibodies was similar to the profile demonstrated by BMA031.
  • FIG. 14 IFN-gamma release from T cells in IVE assay.
  • CD8+ T cells were treated with anti- ⁇ TCR antibodies at various concentrations (see FIG. 2 , x-axis) and co-cultured with autologous dendritic cells pulsed with the CMV peptide 495-503 (pp65) for seven days in an in vitro education (IVE) assay. IFN-gamma release was measured in this assay.
  • FIG. 15 Activation-induced apoptosis by anti- ⁇ TCR antibodies.
  • Antigen stimulated CD8+ T cells were induced to apoptosis by binding of anti- ⁇ TCR antibodies BMA031 and HEBE1 H66.
  • the ability of HEBE1 H66 to induce apoptosis was increased compared to BMA031.
  • FIG. 16 Isolation of glycosylation mutants and aglycosylated antibodies
  • FIG. 17 Binding of ⁇ TCR antibody mutants to human Fc ⁇ RIIIa using Biacore.
  • Biacore was used to assess binding to recombinant human Fc ⁇ RIIIa (V158 & F158).
  • FIG. 18 Binding of ⁇ TCR antibody mutants to human Fc ⁇ RI using Biacore.
  • Biacore was used to assess binding to recombinant human and Fc ⁇ RI.
  • FIG. 19 Cytokine release from PBMCs in presence of glycosylation mutant anti- ⁇ TCR antibodies (day 2).
  • Cytokine release profile for TNFa, GM-CSF, IFNy and MO of anti- ⁇ TCR antibodies was similar to the profile demonstrated by BMA031 and H66 delta AB.
  • FIG. 20 Cytokine release from PBMCs in presence of glycosylation mutant anti- ⁇ TCR antibodies (day 2).
  • Cytokine release profile for IL6, IL4 and IL2 of anti- ⁇ TCR antibodies was similar to the profile demonstrated by BMA031 and H66 delta AB.
  • FIG. 21 Cytokine release from PBMCs in presence of glycosylation mutant anti- ⁇ TCR antibodies (day 4).
  • Cytokine release profile for TNFa, GM-CSF, IFNy and IL10 of anti- ⁇ TCR antibodies was similar to the profile demonstrated by BMA031 and H66 delta AB.
  • FIG. 22 Cytokine release from PBMCs in presence of glycosylation mutant anti- ⁇ TCR antibodies (day 4).
  • Cytokine release profile for IL6, IL4 and IL2 of anti- ⁇ TCR antibodies was similar to the profile demonstrated by BMA031 and H66 delta AB.
  • FIG. 23 Binding profiles of TRACERS.
  • FIG. 24 Cytotoxic activity of different T cell recruitment arms.
  • a panel of humanised BMA031 antibodies have been created and from this panel a number of antibodies have been selected which display cytotoxic activity against tumor antigen expressing cell lines
  • FIG. 25 Cytokine release profile of different T cell recruitment arms.
  • a panel of TRACERs with different T cell recruitment arms show similar cytokine release profiles. Large amounts of cytokines are detected following activation of T cells in the presence of target cells whereas in the presence of only unstimulated human PBMC observed cytokine levels are significantly lower.
  • FIG. 26 Binding of CD52 antibody mutants to human Fc ⁇ RIIIa using Biacore.
  • Biacore was used to assess binding of modified anti-CD52 to recombinant human Fc ⁇ RIIIa (V158).
  • Anti-CD52 comprising S298N/Y300S mutations in the Fc domain were used to assess the effector function of the modified molecule.
  • a humanized monoclonal antibody is an antibody which is composed of a human antibody framework, into which have been grafted complementarity determining regions (CDRs) from a non-human antibody. Changes in the human acceptor framework may also be made.
  • CDRs complementarity determining regions
  • Procedures for the design and production of humanized antibodies are well known in the art, and have been described, for example, in Cabilly et al., U.S. Pat. No. 4,816,567; Cabilly et al., European Patent Application 0 125 023; Boss et al., U.S. Pat. No. 4,816,397; Boss et al., European Patent Application 0 120 694; Neuberger, M. S.
  • antibody unless indicated otherwise, is used to refer to entire antibodies as well as antigen-binding fragments of such antibodies.
  • the term encompasses four-chain IgG molecules, as well as antibody fragments.
  • antibody fragments refers to portions of an intact full-length antibody, for example, as further described below.
  • Antibodies may be of any class, such as IgG, IgA or IgM; and of any subclass, such as IgG1 or IgG4. Different classes and subclasses of immunoglobulin have different properties, which may be advantageous in different applications. For example, IgG4 antibodies have reduced binding to Fc receptors.
  • the claimed antibody be capable of selectively binding its defined cognate antigen, which is the ⁇ TCR.CD3 complex.
  • the antibodies of the invention bind the ⁇ TCR.CD3 complex expressed on cells.
  • the human ⁇ TCR/CD3 complex is the T cell receptor complex presented on the surface of T cells. See, Kuhns et al., (2006) Immunity 24:133-139. This complex is targeted by the murine monoclonal antibody BMA031 (see, European patent application EP 0 403 156; SEQ ID NOs: 1 and 2).
  • Naturally occurring immunoglobulins have a common core structure in which two identical light chains (about 24 kD) and two identical heavy chains (about 55 or 70 kD) form a tetramer.
  • the amino-terminal portion of each chain is known as the variable (V) region and can be distinguished from the more conserved constant (C) regions of the remainder of each chain.
  • V variable region
  • C C-terminal region
  • Within the variable region of the light chain also called the V L domain
  • J region Within the variable region of the heavy chain (also called the V H domain), there is a D region in addition to the J region.
  • CDRs complementarity determining regions
  • FRs conserved framework regions
  • FR1, FR2, FR3 and FR4 The locations of CDR and FR regions and a numbering system have been defined by Kabat et al. (Kabat, E. A. et al., Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, U.S. Government Printing Office (1991), and updates thereof which may be found online).
  • CDR region boundaries have been further defined by IMGT nomenclature.
  • Variable regions of antibodies according to the described embodiments may be found in SEQ ID NOs: 5-7 and 12-16, and may be obtained by humanizing BMA031, that is, by transferring the CDRs of BMA031 to a human framework.
  • Two series of humanized antibodies are described; the HEBE1 series, comprising SEQ ID NOs: 5-7, 12 and 13, and the GL1BM series, comprising heavy chain variable regions as shown in SEQ ID NOs: 8, 15 and 16. In both cases, the light chain variable region used is as shown in SEQ ID NO: 14 (GL1BM VK43).
  • the human frameworks used are IGH3-23 in the case of HEBE1, and IGHV1-3*01 and IGKV3-11*01 in the case of GL1 BM.
  • Constant regions may be derived from any human antibody constant regions. Variable region genes may be cloned into expression vectors in frame with constant region genes to express heavy and light immunoglobulin chains. Such expression vectors can be transfected into antibody producing host cells for antibody synthesis.
  • Human antibody variable and constant regions may be derived from sequence databases.
  • immunoglobulin sequences are available in the IMGT/LIGM database (Giudicelli et al., (2006) Nucleic Acids Res. 34: (suppl. 1):D781-D784) or VBase (vbase.mrc-cpe.cam.ac.uk).
  • Aglycosylated antibodies can have extensively modified functionality; see, Boyd et al. (1996) Mol. Immunol. 32:1311-1318.
  • a “delta ab” or ⁇ ab modification, referred to herein, is an Fc modification as described in Armour et al., (1999) Eur. J. Immunol. 29:2613-2624.
  • Techniques for modifying glycosylation of antibody Fc regions are known in the art, and include chemical, enzymatic and mutational means, for example, mutation of the N297 position in the CH 2 domain. Techniques for mutating antibody genes for producing aglycosylated IgG molecules are described in Tao and Morrison (1989) J. Immunol. 143:2595-2601.
  • Nucleic acids as referred to herein include DNA molecules which encode the antibodies of the invention.
  • Preferred are expression vectors, which are suitable for expressing the antibody genes in a host cell.
  • Expression vectors and host cells for antibody gene expression are known in the art; see, for example, Morrow, K. J. Genetic Engineering & Biotechnology News (Jun. 15, 2008) 28(12), and Backliwal, G. et al. (2008) Nucleic Acids Res. 36(15):e96-e96.
  • the invention encompasses antigen-binding fragments of the humanized anti- ⁇ TCR antibodies.
  • Fragments of the antibodies are capable of binding the ⁇ TCR.CD3 complex. They encompass Fab, Fab′, F(ab′) 2 , and F(v) fragments, or the individual light or heavy chain variable regions or portion thereof. Fragments include, for example, Fab, Fab′, F(ab′) 2 , Fv and scFv. These fragments lack the Fc portion of an intact antibody, clear more rapidly from the circulation, and can have less non-specific tissue binding than an intact antibody. These fragments can be produced from intact antibodies using well known methods, for example by proteolytic cleavage with enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab′) 2 fragments).
  • the antibodies and fragments also encompass single-chain antibody fragments (scFv) that bind to the ⁇ TCR.CD3 complex.
  • An scFv comprises an antibody heavy chain variable region (V H ) operably linked to an antibody light chain variable region (V L ) wherein the heavy chain variable region and the light chain variable region, together or individually, form a binding site that binds ⁇ TCR.
  • An scFv may comprise a V H region at the amino-terminal end and a V L region at the carboxy-terminal end.
  • scFv may comprise a V L region at the amino-terminal end and a V H region at the carboxy-terminal end.
  • V L and V H are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the V L and V H regions pair to form monovalent molecules (known as single chain Fv (scFv)).
  • An scFv may optionally further comprise a polypeptide linker between the heavy chain variable region and the light chain variable region.
  • the antibodies and fragments also encompass domain antibody (dAb) fragments as described in Ward, E. S. et al. (1989) Nature 341:544-546 which consist of a V H domain.
  • dAb domain antibody
  • antibodies and fragments also encompass heavy chain antibodies (HCAb). These antibodies are reported to form antigen-binding regions using only heavy chain variable region, in that these functional antibodies are dimers of heavy chains only (referred to as “heavy-chain antibodies” or “HCAbs”). Accordingly, antibodies and fragments may be heavy chain antibodies (HCAb) that specifically bind to the ⁇ TCR.CD3 complex.
  • HCAb heavy chain antibodies
  • the antibodies and fragments also encompass antibodies that are SMIPs or binding domain immunoglobulin fusion proteins specific for ⁇ TCR.CD3 complex. These constructs are single-chain polypeptides comprising antigen-binding domains fused to immunoglobulin domains necessary to carry out antibody effector functions (see, WO 2005/017148).
  • the antibodies and fragments also encompass diabodies. These are bivalent antibodies in which V H and V L domains are expressed on a single polypeptide chain, but using a linker that is too short to allow for pairing between the two domains on the same chain. This forces the domains to pair with complementary domains of another chain and thereby creates two antigen-binding sites (see, for example, WO 93/11161). Diabodies can be bispecific or monospecific.
  • the antibody or antibody fragment thereof does not cross-react with any target other than the ⁇ TCR.CD3 complex.
  • the antibody or fragment thereof may be modified in order to increase its serum half-life, for example, by adding molecules—such as PEG or other water soluble polymers, including polysaccharide polymers to increase the half-life.
  • bispecific antibodies may resemble single antibodies (or antibody fragments) but have two different antigen binding sites (variable regions).
  • Bispecific antibodies can be produced by various methods—such as chemical techniques, “polydoma” techniques or recombinant DNA techniques.
  • Bispecific antibodies may have binding specificities for at least two different epitopes, at least one of which is the ⁇ TCR.CD3 complex.
  • the other specificity may be selected from any useful or desired specificities including, for example, specificity for human serum albumin for the extension of half-life in vivo.
  • bi-specific antibodies in the clinic for oncology applications is now becoming reality with the tri-functional Catumaxomab (Removmab®) approved for use in cases of malignant ascites and the bi-specific antibody Blinatumomab now in phase II trials in hematological malignancies.
  • These molecules have in common a binding arm which binds to T cells and a second arm which binds to the tumor target cell which results in T cell mediated lysis of the tumor target.
  • these molecules recruit T cells via the CD3 protein located on the cell surface.
  • An alternative to recruitment via CD3 is to make use of the ⁇ T cell receptor ( ⁇ TCR) which is also expressed on the surface of the cell.
  • ⁇ T cell receptor ⁇ T cell receptor
  • Antibody production can be performed by any technique known in the art, including in transgenic organisms such as goats (see, Pollock et al. (1999) J. Immunol. Methods 231:147-157), chickens (see, Morrow, K. J. J. (2000) Genet. Eng. News 20:1-55), mice (see Pollock et al., supra) or plants (see, Doran, P. M. (2000) Curr. Opinion Biotechnol. 11:199-204, Ma. J. K-C. (1998) Nat. Med. 4:601-606, Baez, J. et al. (2000) BioPharm. 13:50-54, Stoger, E. et al. (2000) Plant Mol. Biol. 42:583-590). Antibodies may also be produced by chemical synthesis or by expression of genes encoding the antibodies in host cells.
  • a polynucleotide encoding the antibody is isolated and inserted into a replicable construct or vector such as a plasmid for further propagation or expression in a host cell.
  • Constructs or vectors suitable for the expression of a humanized immunoglobulin according to the described embodiments are available in the art.
  • a variety of vectors are available, including vectors which are maintained in single copy or multiple copies in a host cell, or which become integrated into the host cell's chromosome(s).
  • the constructs or vectors can be introduced into a suitable host cell, and cells which express a humanized immunoglobulin can be produced and maintained in culture.
  • a single vector or multiple vectors can be used for the expression of a humanized immunoglobulin.
  • Polynucleotides encoding the antibody are readily isolated and sequenced using conventional procedures (e.g., oligonucleotide probes).
  • Vectors that may be used include plasmid, virus, phage, transposons, minichromosomes of which plasmids are a typical embodiment.
  • Such vectors further include a signal sequence, origin of replication, one or more marker genes, an enhancer element, a promoter and transcription termination sequences operably linked to the light and/or heavy chain polynucleotide so as to facilitate expression.
  • Polynucleotides encoding the light and heavy chains may be inserted into separate vectors and introduced (e.g., by transformation, transfection, electroporation or transduction) into the same host cell concurrently or sequentially or, if desired, both the heavy chain and light chain can be inserted into the same vector prior to such introduction.
  • a promoter can be provided for expression in a suitable host cell. Promoters can be constitutive or inducible. For example, a promoter can be operably linked to a nucleic acid encoding a humanized immunoglobulin or immunoglobulin chain, such that it directs expression of the encoded polypeptide.
  • suitable promoters for prokaryotic and eukaryotic hosts are available. Prokaryotic promoters include lac, tac, T3, T7 promoters for E.
  • 3-phosphoglycerate kinase or other glycolytic enzymes e.g., enolase, glyceralderhyde 3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose 6 phosphate isomerase, 3-phosphoglycerate mutase and glucokinase.
  • Eukaryotic promoters include inducible yeast promoters such as alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, metallothionein and enzymes responsible for nitrogen metabolism or maltose/galactose utilization; RNA polymerase II promoters including viral promoters such as polyoma, fowlpox and adenoviruses (e.g., adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus (in particular, the immediate early gene promoter), retrovirus, hepatitis B virus, actin, rous sarcoma virus (RSV) promoter and the early or late Simian virus 40 and non-viral promoters such as EF-1 alpha (Mizushima and Nagata (1990) Nucleic Acids Res. 18(17):5322). Those of skill in the art will be able to select the appropriate promoter for expressing a
  • enhancer elements can be included instead of or as well as those found located in the promoters described above.
  • suitable mammalian enhancer sequences include enhancer elements from globin, elastase, albumin, fetoprotein, metallothionine and insulin.
  • an enhancer element from a eukaryotic cell virus such as SV40 enhancer, cytomegalovirus early promoter enhancer, polyoma enhancer, baculoviral enhancer or murine IgG2a locus (see, WO 04/009823).
  • enhancers are often located on the vector at a site upstream to the promoter, they can also be located elsewhere e.g., within the untranslated region or downstream of the polyadenylation signal. The choice and positioning of enhancer may be based upon compatibility with the host cell used for expression.
  • the vectors may comprise a selectable marker for selection of host cells carrying the vector, and, in the case of a replicable vector, an origin of replication.
  • Genes encoding products which confer antibiotic or drug resistance are common selectable markers and may be used in prokaryotic (e.g., f3-lactamase gene (ampicillin resistance), Tet gene (tetracycline resistance) and eukaryotic cells (e.g., neomycin (G418 or geneticin), gpt (mycophenolic acid), ampicillin, or hygromycin resistance genes).
  • Dihydrofolate reductase marker genes permit selection with methotrexate in a variety of hosts.
  • auxotrophic markers of the host e.g., LEU2, URA3, HIS3
  • vectors which are capable of integrating into the genome of the host cell such as retroviral vectors, are also contemplated.
  • polyadenylation and termination signals are operably linked to polynucleotide encoding the antibody of the invention. Such signals are typically placed 3′ of the open reading frame.
  • non-limiting examples of polyadenylation/termination signals include those derived from growth hormones, elongation factor-1 alpha and viral (e.g., SV40) genes or retroviral long terminal repeats.
  • non-limiting examples of polydenylation/termination signals include those derived from the phosphoglycerate kinase (PGK) and the alcohol dehydrogenase 1 (ADH) genes.
  • PGK phosphoglycerate kinase
  • ADH alcohol dehydrogenase 1
  • polyadenylation signals are typically not required and it is instead usual to employ shorter and more defined terminator sequences.
  • the choice of polyadenylation/termination sequences may be based upon compatibility with the host cell used for expression.
  • other features that can be employed to enhance yields include chromatin remodeling elements, introns and host cell specific codon modification.
  • the codon usage of the antibodies of the invention can be modified to accommodate codon bias of the host cell such to augment transcript and/or product yield (e.g., Hoekema, A. et al. (1987) Mol. Cell Biol. 7(8):2914-24).
  • the choice of codons may be based upon compatibility with the host cell used for expression.
  • the invention thus relates to isolated nucleic acid molecules that encode the humanized immunoglobulins, or heavy or light chains, thereof.
  • the invention also relates to isolated nucleic acid molecules that encode an antigen-binding portion of the immunoglobulins and their chains.
  • the antibodies can be produced, for example, by the expression of one or more recombinant nucleic acids encoding the antibody in a suitable host cell.
  • the host cell can be produced using any suitable method.
  • the expression constructs e.g., one or more vectors, e.g., a mammalian cell expression vector
  • the resulting cell can be maintained (e.g., in culture, in an animal, in a plant) under conditions suitable for expression of the construct(s) or vector(s).
  • Host cells can be prokaryotic, including bacterial cells such as E.
  • coli e.g., strain DH5aTM
  • PerC6 Cell, Leiden, NL
  • B. subtilis and/or other suitable bacteria eukaryotic cells, such as fungal or yeast cells (e.g., Pichia pastoris, Aspergillus sp., Saccharomyces cerevisiae, Schizosaccharomyces pombe, Neurospora crassa ), or other lower eukaryotic cells, and cells of higher eukaryotes such as those from insects (e.g., Drosophila Schnieder S2 cells, Sf9 insect cells) (WO 94/126087 (O'Connor)), BTI-TN-5B1-4 (High FiveTM) insect cells (Invitrogen), mammals (e.g., COS cells, such as COS-1 (ATCC Accession No.
  • CRL-1650 and COS-7 (ATCC Accession No. CRL-1651), CHO (e.g., ATCC Accession No. CRL-9096), CHO DG44 (Urlaub, G. and Chasin, L. A. (1980) Proc. Natl. Acad. Sci. USA, 77(7):4216-4220), 293 (ATCC Accession No. CRL-1573), HeLa (ATCC Accession No. CCL-2), CVI (ATCC Accession No. CCL-70), WOP (Dailey, L., et al. (1985) J. Virol., 54:739-749), 3T3, 293T (Pear, W. S., et al. (1993) Proc. Natl.
  • the host cell is not part of a multicellular organism (e.g., plant or animal), e.g., it is an isolated host cell or is part of a cell culture.
  • Host cells may be cultured in spinner flasks, shake flasks, roller bottles, wave reactors (e.g., System 1000 from wavebiotech.com) or hollow fibre systems but it is preferred for large scale production that stirred tank reactors or bag reactors (e.g., Wave Biotech, Somerset, N.J. USA) are used particularly for suspension cultures.
  • Stirred tank reactors can be adapted for aeration using e.g., spargers, baffles or low shear impellers. For bubble columns and airlift reactors, direct aeration with air or oxygen bubbles maybe used.
  • the medium can be supplemented with a cell protective agent such as pluronic F-68 to help prevent cell damage as a result of the aeration process.
  • a cell protective agent such as pluronic F-68 to help prevent cell damage as a result of the aeration process.
  • microcarriers maybe used as growth substrates for anchorage dependent cell lines, or the cells maybe adapted to suspension culture.
  • the culturing of host cells, particularly vertebrate host cells may utilize a variety of operational modes such as batch, fed-batch, repeated batch processing (see, Drapeau et al. (1994) Cytotechnology 15:103-109), extended batch process or perfusion culture.
  • recombinantly transformed mammalian host cells may be cultured in serum-containing media such media comprising fetal calf serum (FCS), it is preferred that such host cells are cultured in serum-free media such as disclosed in Keen et al. (1995) Cytotechnology 17:153-163, or commercially available media such as ProCHO-CDM or UltraCHOTM (Cambrex N.J., USA), supplemented where necessary with an energy source such as glucose and synthetic growth factors such as recombinant insulin.
  • FCS fetal calf serum
  • serum-free media such as disclosed in Keen et al. (1995) Cytotechnology 17:153-163, or commercially available media such as ProCHO-CDM or UltraCHOTM (Cambrex N.J., USA), supplemented where necessary with an energy source such as glucose and synthetic growth factors such as recombinant insulin.
  • the serum-free culturing of host cells may require that those cells are adapted to grow in serum-free conditions.
  • One adaptation approach is to culture such host cells in serum containing media and repeatedly exchange 80% of the culture medium for the serum-free media so that the host cells learn to adapt in serum-free conditions (see, e.g., Scharfenberg, K. et al. (1995) Animal Cell Technology: Developments Towards the 21 st Century (Beuvery, E. C. et al., eds), pp.619-623, Kluwer Academic publishers).
  • Antibodies according to the described embodiments may be secreted into the medium and recovered and purified therefrom using a variety of techniques to provide a degree of purification suitable for the intended use.
  • the use of therapeutic antibodies for the treatment of human patients typically mandates at least 95% purity as determined by reducing SDS-PAGE, more typically 98% or 99% purity, when compared to the culture media comprising the therapeutic antibodies.
  • cell debris from the culture media can be removed using centrifugation followed by a clarification step of the supernatant using e.g., microfiltration, ultrafiltration and/or depth filtration.
  • the antibody can be harvested by microfiltration, ultrafiltration or depth filtration without prior centrifugation.
  • HA hydroxyapatite
  • affinity chromatography optionally involving an affinity tagging system such as polyhistidine
  • HIC hydrophobic interaction chromatography
  • the antibodies following various clarification steps, are captured using Protein A or G affinity chromatography followed by further chromatography steps such as ion exchange and/or HA chromatography, anion or cation exchange, size exclusion chromatography and ammonium sulphate precipitation.
  • virus removal steps may also be employed (e.g., nanofiltration using, e.g., a DV-20 filter).
  • a purified preparation comprising at least 10 mg/ml or greater, e.g., 100 mg/ml or greater of the antibody of the invention is provided and, therefore, forms another embodiment of the invention. Concentration to 100 mg/ml or greater can be generated by ultracentrifugation. Such preparations are substantially free of aggregated forms of antibodies of the invention.
  • Bacterial systems are particularly suited for the expression of antibody fragments. Such fragments are localized intracellularly or within the periplasm. Insoluble periplasmic proteins can be extracted and refolded to form active proteins according to methods known to those skilled in the art, see, Sanchez et al. (1999) J. Biotechnol. 72:13-20; Cupit, P. M. et al. (1999) Lett. Appl. Microbiol. 29:273-277.
  • the present invention also relates to cells comprising a nucleic acid, e.g., a vector, of the invention (e.g., an expression vector).
  • a nucleic acid i.e., one or more nucleic acids
  • a construct i.e., one or more constructs, e.g., one or more vectors
  • nucleic acid(s) can be introduced into a suitable host cell by a method appropriate to the host cell selected (e.g., transformation, transfection, electroporation, infection), with the nucleic acid(s) being, or becoming, operably linked to one or more expression control elements (e.g., in a vector, in a construct created by processes in the cell, integrated into the host cell genome).
  • Host cells can be maintained under conditions suitable for expression (e.g., in the presence of inducer, suitable media supplemented with appropriate salts, growth factors, antibiotic, nutritional supplements, etc.), whereby the encoded polypeptide(s) are produced.
  • the encoded humanized antibody can be isolated, for example, from the host cells, culture medium, or milk. This process encompasses expression in a host cell (e.g., a mammary gland cell) of a transgenic animal or plant (e.g., tobacco) (see, e.g., WO 92/03918).
  • T cell activity is desirable in a number of situations in which immunosuppression is warranted, and/or an autoimmune condition occurs. Accordingly, targeting of the ⁇ TCR.CD3 complex is indicated in the treatment of diseases involving an inappropriate or undesired immune response, such as inflammation, autoimmunity, and other conditions involving such mechanisms. In one embodiment, such disease or disorder is an autoimmune and/or inflammatory disease.
  • autoimmune and/or inflammatory diseases are Systemic Lupus Erythematosus (SLE), Rheumatoid Arthritis (RA) and inflammatory bowel disease (IBD) (including ulcerative colitis (UC) and Crohn's disease (CD)), multiple sclerosis (MS), scleroderma and type 1 diabetes (T1D), and other diseases and disorders, such as PV (pemphigus vulgaris), psoriasis, atopic dermatitis, celiac disease, Chronic Obstructive Lung disease, Hashimoto's thyroiditis, Graves' disease (thyroid), Sjogren's syndrome, Guillain-barré syndrome, Goodpasture's syndrome, Addison's disease, Wegener's granulomatosis, primary biliary sclerosis, sclerosing cholangitis, autoimmune hepatitis, polymyalgia rheumatica, Raynaud's phenomenon, temporal arteritis,
  • such disease or disorder is SLE, RA or IBD. In one embodiment, such disease or disorder is MS.
  • the antibodies according to the described embodiments are used to aid transplantation by immunosuppressing the subject. Such use alleviates graft-versus-host disease.
  • graft-versus-host disease For a description of existing treatments for graft-versus-host disease, see, e.g., Svennilson, Bone Marrow Transplantation (2005) 35:S65-S67, and references cited therein.
  • the antibodies of the invention may be used in combination with other available therapies.
  • combination therapy may include administration of an antibody of the present invention together with a medicament, which together with the antibody comprises an effective amount for preventing or treating such autoimmune diseases.
  • the combination therapy may encompass one or more of an agent that promotes the growth of pancreatic beta-cells or enhances beta-cell transplantation, such as beta cell growth or survival factors or immunomodulatory antibodies.
  • said combination therapy may encompass one or more of methotrexate, an anti-TNF- ⁇ antibody, a TNF- ⁇ receptor-Ig fusion protein, an anti-IL-15 or anti-IL-21 antibody, a non-steroidal anti-inflammatory drug (NSAID), or a disease-modifying anti-rheumatic drug (DMARD).
  • the additional agent may be a biological agent such as an anti-TNF agent (e.g., Enbrel®, infliximab (Remicade®) and adalimumab (Humira®) or rituximab (Rituxan®).
  • hematopoietic growth factor(s) such as erythropoietin, G-CSF, GM-CSF, IL-3, IL-11, thrombopoietin, etc.
  • antimicrobial(s) such as antibiotic, antiviral, antifungal drugs
  • the additional agent may be one or more of tar and derivatives thereof, phototherapy, corticosteroids, Cyclosporine A, vitamin D analogs, methotrexate, p38 mitogen-activated protein kinase (MAPK) inhibitors, as well as biologic agents such as anti-TNF-agents and Rituxan®.
  • the additional agent may be one or more of aminosalicylates, corticosteroids, immunomodulators, antibiotics, or biologic agents such as Remicade® and Humira®.
  • combination treatment may be carried out in any way as deemed necessary or convenient by the person skilled in the art and for the purpose of this specification, no limitations with regard to the order, amount, repetition or relative amount of the compounds to be used in combination is contemplated. Accordingly, the antibodies according to the described embodiments may be formulated into pharmaceutical compositions for use in therapy.
  • a pharmaceutical composition comprising an antibody according to the invention, or a ligand or ligands identifiable by an assay method as defined in the previous aspect of the invention.
  • Ligands may be immunoglobulins, peptides, nucleic acids or small molecules, as discussed herein. They are referred to, in the following discussion, as “compounds”.
  • a pharmaceutical composition according to the invention is a composition of matter comprising a compound or compounds capable of modulating T cell activity as an active ingredient.
  • the compound is in the form of any pharmaceutically acceptable salt, or e.g., where appropriate, an analog, free base form, tautomer, enantiomer racemate, or combination thereof.
  • the active ingredients of a pharmaceutical composition comprising the active ingredient according to the invention are contemplated to exhibit therapeutic activity, for example, in the treatment of graft-versus-host disease, when administered in amount which depends on the particular case.
  • one or more compounds of the invention may be used in combination with any art recognized compound known to be suitable for treating the particular indication in treating any of the aforementioned conditions. Accordingly, one or more compounds of the invention may be combined with one or more art recognized compounds known to be suitable for treating the foregoing indications such that a convenient, single composition can be administered to the subject. Dosage procedures may be adjusted to provide the optimum therapeutic response.
  • doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.
  • the active ingredient may be administered in a convenient manner such as by the oral, intravenous (where water soluble), intramuscular, subcutaneous, intranasal, intradermal or suppository routes or implanting (e.g., using slow release molecules).
  • the active ingredient may be required to be coated in a material to protect said ingredients from the action of enzymes, acids and other natural conditions which may inactivate said ingredient.
  • the active ingredient in order to administer the active ingredient by means other than parenteral administration, it will be coated by, or administered with, a material to prevent its inactivation.
  • the active ingredient may be administered in an adjuvant, co-administered with enzyme inhibitors or in liposomes.
  • Adjuvant is used in its broadest sense and includes any immune stimulating compound such as interferon.
  • Adjuvants contemplated herein include resorcinols, non-ionic surfactants such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether.
  • Enzyme inhibitors include pancreatic trypsin.
  • Liposomes include water-in-oil-in-water CGF emulsions as well as conventional liposomes.
  • the active ingredient may also be administered parenterally or intraperitoneally.
  • Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • the pharmaceutical forms suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
  • the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of superfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thirmerosal, and the like.
  • isotonic agents for example, sugars or sodium chloride.
  • Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin.
  • Sterile injectable solutions are prepared by incorporating the active ingredient in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the sterilized active ingredient into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above.
  • the preferred methods of preparation are vacuum drying and the freeze-drying technique which yield a powder of the active ingredient plus any additional desired ingredient from previously sterile-filtered solution thereof.
  • any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed.
  • the active ingredient may be incorporated into sustained-release preparations and formulations.
  • pharmaceutically acceptable carrier and/or diluent includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like.
  • the use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, use thereof in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
  • Dosage unit form refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • the specification for the novel dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such as active material for the treatment of disease in living subjects having a diseased condition in which bodily health is impaired.
  • compositions containing supplementary active ingredients are compounded for convenient and effective administration in effective amounts with a suitable pharmaceutically acceptable carrier in dosage unit form.
  • dosages are determined by reference to the usual dose and manner of administration of the said ingredients.
  • peptides may be modified in order to improve their ability to cross a cell membrane.
  • U.S. Pat. No. 5,149,782 discloses the use of fusogenic peptides, ion-channel forming peptides, membrane peptides, long-chain fatty acids and other membrane blending agents to increase protein transport across the cell membrane.
  • a method for treating a condition associated with an aberrant immune response comprising administering to a subject a therapeutically effective amount of a ligand identifiable using an assay method as described above.
  • CD8+ T cells were treated with anti- ⁇ TCR antibodies at various concentrations (see, FIG. 2 , x-axis) and co-cultured with autologous dendritic cells (DCs) pulsed with the CMV peptide 495-503 (pp65) for seven days in an in vitro education (IVE) assay.
  • DCs autologous dendritic cells
  • PBMC Normal donor aphaeresis products from HLA-A2 + individuals were obtained from HemaCare Corp., Van Nuys, Calif.). PBMC were isolated by centrifugation over Ficoll (GE Healthcare, Piscataway, N.J.). CD8+ T cells were isolated using magnetic beads (Invitrogen, Carlsbad, Calif.) according to the manufacturer's instructions. To generate autologous immature dendritic cells, PBMC were resuspended in RPMI 1640/5% human AB serum (Sigma), plated in triple flasks (Corning) and incubated for more than 2 hours at 37° C./5% CO 2 .
  • the adherent monocytes were then rinsed with PBS and cultured for 6 days in RPMI 1640/5% human AB serum supplemented with GM-CSF (Immunex, Seattle, Wash.) and IL-4 (PeproTech, Rocky Hill, N.J.). Prior to establishing the T cell/DC co-cultures, the DCs were pulsed with peptides (10 ug/ml) for 4 hours and then matured.
  • Mature dendritic cells were generated by the addition of 50 ng/ml TNF-alpha, 25 ng/ml IL-1 ⁇ , 10 ng/ml IL-6, 500 ng/ml PGE-2 (PeproTech, Rocky Hill, N.J.) and culturing the dendritic cells for an additional 24 hours.
  • the peptide-pulsed DCs were then added to the previously isolated CD8+ T cells at a T:DC ratio of 10:1.
  • Cultures were immediately fed with IL-2 (100 IU/ml) added to the cultures.
  • the cultures were supplemented with IL-2 (100 IU/ml) on day 4.
  • the bulk cultures were assayed for peptide reactivity in a chromium release assay on day 7.
  • the graph in FIG. 2 shows lysis data from the chromium release assay, where untreated T cells were successfully educated against pp65 peptide and able to lyse specific targets at >50%.
  • BMA031 inhibited education of these T cells, as they were unable to lyse specific targets in a dose-dependent manner.
  • Humanized antibody EuCIV3 was less potent than BMA031 and was only able to inhibit education at the highest dose.
  • PBMC proliferation assay human PBMC were cultured with increasing concentrations of therapeutic antibody for 72 hours, 3 H-thymidine was added and cells were harvested 18 hours later.
  • human PBMC were cultured with increasing concentrations of therapeutic antibody and were analyzed daily for cell counts and viability (Vi-Cell, Beckman Coulter) out to day 7. Cell supernatants were also harvested, stored at ⁇ 20° C. and analyzed on an 8-plex cytokine panel (Bio-Rad).
  • BMA031 does not induce: (i) PBMC proliferation; (ii) T cell depletion; (iii) CD25 expression; or (iv) cytokine release.
  • OKT3 does induce all of the aforementioned effects.
  • BMA031 and OKT3 are capable of blocking the education of CD8+ cells to peptide in an in vitro education (IVE) assay and are also capable of blocking a mixed lymphocyte reaction (MLR).
  • BMA031 also induces apoptosis of activated T cells (activation-induced cell death; AICD).
  • HuIgG1 delta ab and HuIgG4 agly anti- ⁇ TCR antibodies had the same in vitro profile as BMA031 MoIgG2b (Table 1).
  • the strategy for optimization of the humanized antibodies was based upon mutagenesis and functional screening. Optimization was started with block changes of amino acid residues in one of each of the four framework regions of the variable domains, from mouse to human. Key framework regions were identified in each of the GL1BM HC, GL1BM LC and HEBE1 HC series. Following this identification, individual residues within those framework regions were mutated to human germline residues from the original mouse sequence. Framework residues for which identity with the mouse sequence was found to be important to retaining the binding properties of the antibody were retained as mouse residues. Otherwise, framework residues were changed to match the human germline amino acid sequence. This was continued across the sequence until the minimal number of mouse residues, to retain the original binding properties of the antibody, were identified. See FIG. 5 . We have demonstrated that several of the antibodies from these series have an improved binding compared to BMA031 as determined by antibody off-rate from T cells ( FIGS. 6, 7 and 8 ).
  • the antibodies are active in preventing the immune response in an IVE ( FIGS. 9, 10 and 11 ) and a MLR assay.
  • IVE assay tetramer binding was used as a quantitative measurement for the IVE.
  • the percentage of cells which were antigen specific was determined by staining the T cells with a directly labeled tetramer that is specific for the educating peptide. Briefly, day 7 CD8+ T cells from the IVE were stained with tetramer by standard flow cytometry staining protocols and analyzed on BD FACSCalibur.
  • the humanized antibodies demonstrated comparable levels of proliferative potential on PBMCs and cytokine release as compared to BMA031 ( FIGS. 12 and 13 ).
  • the antibodies also showed an ability to inhibit the release of IFN ⁇ from T cells in an IVE assay ( FIG. 14 ).
  • a number of these antibodies have an improved ability to elicit activation-induced cell death (AICD) of activated ⁇ TCR-positive T cells compared to BMA031 ( FIG. 15 ).
  • AICD activation-induced cell death
  • antigen-specific CD8+ T cells were cultured with therapeutic antibody. At 24 hours, 48 hours and 72 hours cells were stained for apoptosis markers Annexin-V and 7-AAD. Cells were also stained with tetramer to track apoptosis with effects on antigen-specific T cells.
  • Engineered Fc variants was designed and generated where a glycosylation site is introduced at amino acid Ser 298 position, next to the naturally-occurring Asn297 site.
  • the glycosylation at Asn297 was either kept or knocked out by mutations. Mutations and glycosylation results are set forth in Table 2.
  • HEBE1 Agly IgG4 and HEBE1 ⁇ ab IgG1 in pCEP4 were used as controls in HEK293 transfection.
  • the mutants, wt and controls were transfected into HEK293-EBNA cells in triple-flask for expression. Proteins were purified from 160 ml of conditioned media (CM) with 1 ml HiTrap protein A columns (GE) on multichannel peristaltic pump. Five micrograms of each supernatant were analyzed on 4-20% Tris-Glycine reducing and non-reducing SDS-PAGE (see FIG. 16 ). The heavy chain of the aglycosylated mutants (N297Q, T299A, and Agly control, is lower (arrow in black), consistent with the loss of the glycans in these antibody.
  • CM conditioned media
  • GE HiTrap protein A columns
  • the heavy chains of the engineered glycosylated antibodies migrate the same way as the wild-type control. This result is consistent with the expected outcome of engineered glycosylation site at 298 positions. SEC-HPLC analysis indicated that all mutants are expressed as monomers.
  • the Engineered H66 IgG1 Fc variants were partially reduced with 20 mM DTT at 37° C. for 30 min.
  • the samples were analyzed by capillary LC/MS on an Agilent 1100 capillary HPLC system coupling with a QSTAR qq TOF hybrid system (Applied Biosystem).
  • Bayesian protein reconstruct with baseline correction and computer modeling in Analyst QS 1.1 (Applied Bisoystem) was used for data analysis.
  • mutant S298N/T299A/Y300S H66 antibody lead one glycosylation site was observed at amino acid 298 with bi-antennary and tri-antennary complex-type glycans detected as the major species, as well as G0F, G1F and G2F.
  • Biacore was used to assess binding to recombinant human Fc ⁇ RIIIa (V158 & F158) and Fc ⁇ RI. All 4 flowcells of a CM5 chip were immobilized with anti-HPC4 antibody via the standard amine coupling procedure provided by Biacore. The anti-HPC4 antibody was diluted to 50 ⁇ g/mL in 10 mM sodium acetate pH 5.0 for the coupling reaction and injected for 25 min at 5 ⁇ L/min. Approximately 12,000 RU of antibody was immobilized to the chip surface.
  • Recombinant human Fc ⁇ RIIIa-V158 and Fc ⁇ RIIIa-F158 were diluted to 0.6 ⁇ g/mL in binding buffer, HBS-P with 1 mM CaCl 2 , and injected to flowcells 2 and 4, respectively, for 3 min at 5 ⁇ L/min to capture 300-400 RU receptor to the anti-HPC4 chip.
  • three times more rhFc ⁇ RIIIa was captured to the anti-HPC4 surface than usually used in this assay.
  • Flowcells 1 and 3 were used as reference controls. Each antibody was diluted to 200 nM in binding buffer and injected over all 4 flowcells for 4 min, followed by 5 min dissociation in buffer. The surfaces were regenerated with 10 mM EDTA in HBS-EP buffer for 3 min at 20 ⁇ L/min.
  • Biacore was also used to compare the Fc ⁇ RI binding.
  • Anti-tetra His antibody was buffer exchanged into 10 mM sodium acetate pH 4.0 using a Zeba Desalting column and diluted to 25 ⁇ g/mL in the acetate buffer for amine coupling.
  • Two flowcells of a CM5 chip were immobilized with ⁇ 9000 RU of the anti-Tetra-His antibody after 20 min injection at 5 ⁇ L/min. Similar to the previous experiment, ten times more Fc ⁇ RI was captured to the anti-tetra-His surface in order to compare weak binders.
  • Recombinant human Fc ⁇ RI was diluted 10 ⁇ g/mL in HBS-EP binding buffer and injected to flowcell 2 for 1 min at 5 ⁇ L/min to capture ⁇ 1000 RU receptor to the anti-tetra-His chip.
  • a single concentration of antibody, 100 nM was injected for 3 min at 30 ⁇ L/min over the captured receptor and control surface. Dissociation was monitored for 3 min. The surface was regeneration with two 30 sec injections of 10 mM glycine pH 2.5 at 20 ⁇ L/min.
  • the stability of the S298N/T299A/Y300S antibody mutant was monitored by a Far-UV CD thermo melting experiment where the CD signal at 216 nm and 222 nm was monitored as temperature increases that eventually leads to the unfolding of the antibody.
  • the CD spectra were collected on a Jasco 815 spectrophotometer at a protein concentration of approximately 0.5 mg/mL in PBS buffer in a quartz cuvette (Hellma, Inc) with a path length of 10 mm. Temperature was controlled by a thermoelectric peltier (Jasco model AWC100) and was ramped at a rate of 1° C./min from 25-89° C. CD signal and HT voltage were both collected.
  • PBMC proliferation and cytokine release assays were conducted as set forth in Example 2. Normal donor PBMC were thawed and treated under the following conditions (all in media containing complement):
  • Cytokines were harvested at day 2 (D2) and day 4 (D4) for Bioplex Analysis (IL2, IL4, IL6, IL8, IL10, GM-CSF, IFNg, TNFa). Cells were stained at D4 for CD4, CD8, CD25 and abTCR expression.
  • H66 S298N/T299A/Y300S behaved similarly to the H66 deltaAB in all cell based assays, showing minimal T-cell activation by CD25 expression; binding to abTCR, although with slightly different kinetics to deltaAB; minimal cytokine release at both D2 and D4 time points; the mutant was in fact superior to deltaAB at D4 in respect of several of the cytokines.
  • the S298N/T299A/Y300S mutant thus eliminated effector function as effectively as the deltaAB mutation.
  • Bi-specific molecules were constructed comprised of two single chain antibodies (scFv) linked together via a short amino acid linker whereby one arm is capable of binding a tumor target and the other capable of binding T cells via the ⁇ TCR.
  • the bispecific molecule is referred to herein as a TRACER (T cell Receptor Activated Cytotoxic EnableR).
  • sequences of the heavy and light chains are set forth in SEQ ID nos 14-16 and 20-24
  • Characterization of these molecules comprised an assessment of binding to tumor target and T cells, in vitro cytotoxic activity and cytokine release profile in the presence and absence of tumor target cells.
  • the binding profile assessed by flow cytometery shows that anti- ⁇ TCR bi-specific antibodies are able to bind both the tumor target cell line and T cells. See FIG. 23 .
  • glycosylation mutant S298N/Y300S was also prepared in an anti-CD52 antibody (clone 2C3) to see whether the effector function modulation with the loss of Fc ⁇ RIII binding applies to a different antibody backbone.
  • S298N/Y300S 2C3 variant DNA was prepared by quick change mutagenesis. The protein was purified from conditioned media after HEK293 transient transfection. Anti-CD52 2C3 wild-type antibody was produced in parallel as a control. Biacore was used to characterize the antigen-binding, Fc ⁇ RIII, and binding properties of the purified antibodies (see FIG. 26 ).
  • the S298N/Y300S 2C3 variant binds to CD52 peptide tightly and the binding sensorgram is undistinguishable with the wild-type control, suggesting that this mutation on the Fc domain does not affect its antigen binding ( FIG. 26A ).
  • Fc ⁇ RIII receptor (Val158) was used in binding studies.
  • the mutant and wild-type control antibody were diluted to 200 nM and injected to HPC4-tag captured Fc ⁇ RIIIa.
  • Fc ⁇ RIII binding is almost undetectable for the S298N/Y300S mutant, which indicates loss of effector function with this variant ( FIG. 26B ).
  • the mutant also binds to FcRn receptor with the same affinity as the wild-type antibody control so we expect no change in its circulation half-life or other pharmacokinetic properties. (see FIG. 26C ).
  • the S298N/Y300S mutation is applicable to antibodies in general, to reduce or eliminate undesired Fc effector function, for example through engagement of human Fc ⁇ receptors.

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Publication number Priority date Publication date Assignee Title
US11186638B2 (en) 2011-09-12 2021-11-30 Genzyme Corporation Anti-αβTCR antibody
US20220098299A1 (en) * 2013-10-30 2022-03-31 Genzyme Corporation Methods for enhancing immunosuppressive therapy by multiple administration of alpha beta tcr-binding polypeptide
US11697690B2 (en) 2014-03-19 2023-07-11 Genzyme Corporation Site-specific glycoengineering of targeting moieties
US11807690B2 (en) 2013-03-11 2023-11-07 Genzyme Corporation Hyperglycosylated binding polypeptides

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PT2895513T (pt) * 2012-09-12 2018-10-08 Genzyme Corp Polipéptidos contendo fc com glicosilação alterada e função efetora reduzida
US9790268B2 (en) 2012-09-12 2017-10-17 Genzyme Corporation Fc containing polypeptides with altered glycosylation and reduced effector function
AU2015330869B2 (en) 2014-10-09 2021-07-08 Genzyme Corporation Glycoengineered antibody drug conjugates
WO2016059220A1 (en) 2014-10-16 2016-04-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Tcr-activating agents for use in the treatment of t-all
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US20190016804A1 (en) 2017-07-14 2019-01-17 Immatics Biotechnologies Gmbh Dual specificity polypeptide molecule
RU2708558C1 (ru) * 2018-12-13 2019-12-09 Российская Федерация, от имени которой выступает ФОНД ПЕРСПЕКТИВНЫХ ИССЛЕДОВАНИЙ Способ создания противоопухолевой иммунологической защиты к клеткам лимфомы EL-4
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MX2022015872A (es) 2020-06-11 2023-05-16 Provention Bio Inc Metodos y composiciones para prevenir diabetes tipo 1.
WO2023227018A1 (zh) * 2022-05-25 2023-11-30 羿尊生物医药（浙江）有限公司 靶向细胞膜受体蛋白的融合蛋白及其应用

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0403156A1 (en) 1989-06-07 1990-12-19 Genzyme Corporation Improved monoclonal antibodies against the human alpha/beta t-cell receptor, their production and use
WO2004099249A2 (en) 2003-05-02 2004-11-18 Xencor, Inc. Optimized fc variants and methods for their generation
WO2006105021A2 (en) 2005-03-25 2006-10-05 Tolerrx, Inc. Gitr binding molecules and uses therefor
US20080038260A1 (en) 2006-06-19 2008-02-14 Tolerrx, Inc. ILT3 binding molecules and uses therefor
WO2008086006A2 (en) 2007-01-09 2008-07-17 Biogen Idec Ma Inc. Sp35 antibodies and uses thereof
WO2010027797A1 (en) 2008-08-26 2010-03-11 Macrogenics Inc. T-cell receptor antibodies and methods of use thereof
US20100190247A1 (en) 2003-12-04 2010-07-29 Xencor, Inc. Methods of Generating Variant Proteins with Increased Host String Content
WO2011109400A2 (en) 2010-03-04 2011-09-09 Macrogenics,Inc. Antibodies reactive with b7-h3, immunologically active fragments thereof and uses thereof

Family Cites Families (114)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB8308235D0 (en) 1983-03-25 1983-05-05 Celltech Ltd Polypeptides
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
GB8422238D0 (en) 1984-09-03 1984-10-10 Neuberger M S Chimeric proteins
US5225539A (en) 1986-03-27 1993-07-06 Medical Research Council Recombinant altered antibodies and methods of making altered antibodies
GB8607679D0 (en) 1986-03-27 1986-04-30 Winter G P Recombinant dna product
WO1988007089A1 (en) 1987-03-18 1988-09-22 Medical Research Council Altered antibodies
US5149782A (en) 1988-08-19 1992-09-22 Tanox Biosystems, Inc. Molecular conjugates containing cell membrane-blending agents
EP0359096B1 (en) 1988-09-15 1997-11-05 The Trustees Of Columbia University In The City Of New York Antibodies having modified carbohydrate content and methods of preparation and use
US6780613B1 (en) 1988-10-28 2004-08-24 Genentech, Inc. Growth hormone variants
US5108921A (en) 1989-04-03 1992-04-28 Purdue Research Foundation Method for enhanced transmembrane transport of exogenous molecules
ATE158021T1 (de) 1990-08-29 1997-09-15 Genpharm Int Produktion und nützung nicht-menschliche transgentiere zur produktion heterologe antikörper
DE69233482T2 (de) 1991-05-17 2006-01-12 Merck & Co., Inc. Verfahren zur Verminderung der Immunogenität der variablen Antikörperdomänen
EP0617706B1 (en) 1991-11-25 2001-10-17 Enzon, Inc. Multivalent antigen-binding proteins
US5622929A (en) 1992-01-23 1997-04-22 Bristol-Myers Squibb Company Thioether conjugates
EP0627932B1 (en) 1992-11-04 2002-05-08 City Of Hope Antibody construct
WO1994026087A2 (en) 1993-05-14 1994-11-24 Connor Kim C O Recombinant protein production and insect cell culture and process
US5429746A (en) 1994-02-22 1995-07-04 Smith Kline Beecham Corporation Antibody purification
US5792456A (en) 1994-08-04 1998-08-11 Bristol-Myers Squibb Company Mutant BR96 antibodies reactive with human carcinomas
ATE208633T1 (de) 1994-09-16 2001-11-15 Merck Patent Gmbh Immunokonjugate
GB9422383D0 (en) 1994-11-05 1995-01-04 Wellcome Found Antibodies
US6096871A (en) 1995-04-14 2000-08-01 Genentech, Inc. Polypeptides altered to contain an epitope from the Fc region of an IgG molecule for increased half-life
US6121022A (en) 1995-04-14 2000-09-19 Genentech, Inc. Altered polypeptides with increased half-life
US5869046A (en) 1995-04-14 1999-02-09 Genentech, Inc. Altered polypeptides with increased half-life
US5739277A (en) 1995-04-14 1998-04-14 Genentech Inc. Altered polypeptides with increased half-life
WO1996040662A2 (en) 1995-06-07 1996-12-19 Cellpro, Incorporated Aminooxy-containing linker compounds and their application in conjugates
US5989830A (en) 1995-10-16 1999-11-23 Unilever Patent Holdings Bv Bifunctional or bivalent antibody fragment analogue
GB9524973D0 (en) 1995-12-06 1996-02-07 Lynxvale Ltd Viral vectors
SE9601245D0 (sv) 1996-03-29 1996-03-29 Pharmacia Ab Chimeric superantigens and their use
UA71889C2 (uk) 1996-04-02 2005-01-17 Йєда Рісерч Енд Дівелопмент Ко. Лтд. Модулятори зв'язаного з рецептором tnf фактора (traf), їх одержання та застосування
DE69738522T2 (de) 1996-08-02 2009-04-02 Bristol-Myers Squibb Co. Ein verfahren zur inhibierung immunglobulininduzierter toxizität aufgrund von der verwendung von immunoglobinen in therapie und in vivo diagnostik
WO1998023289A1 (en) 1996-11-27 1998-06-04 The General Hospital Corporation MODULATION OF IgG BINDING TO FcRn
US6277375B1 (en) 1997-03-03 2001-08-21 Board Of Regents, The University Of Texas System Immunoglobulin-like domains with increased half-lives
ATE419009T1 (de) 1997-10-31 2009-01-15 Genentech Inc Methoden und zusammensetzungen bestehend aus glykoprotein-glykoformen
US6953675B2 (en) 1997-11-06 2005-10-11 Immunomedics, Inc. Landscaped antibodies and antibody fragments for clinical use
US6528624B1 (en) 1998-04-02 2003-03-04 Genentech, Inc. Polypeptide variants
US6194551B1 (en) 1998-04-02 2001-02-27 Genentech, Inc. Polypeptide variants
US6242195B1 (en) 1998-04-02 2001-06-05 Genentech, Inc. Methods for determining binding of an analyte to a receptor
DE69937291T2 (de) 1998-04-02 2008-07-10 Genentech, Inc., South San Francisco Antikörpervarianten und fragmente davon
GB9809951D0 (en) 1998-05-08 1998-07-08 Univ Cambridge Tech Binding molecules
AU770555B2 (en) 1998-08-17 2004-02-26 Abgenix, Inc. Generation of modified molecules with increased serum half-lives
WO2000017226A1 (en) 1998-09-23 2000-03-30 The Regents Of The University Of California Synthetic peptides, conjugation reagents and methods
EP1006183A1 (en) 1998-12-03 2000-06-07 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Recombinant soluble Fc receptors
US7183387B1 (en) 1999-01-15 2007-02-27 Genentech, Inc. Polypeptide variants with altered effector function
US6737056B1 (en) 1999-01-15 2004-05-18 Genentech, Inc. Polypeptide variants with altered effector function
KR20060067983A (ko) 1999-01-15 2006-06-20 제넨테크, 인크. 효과기 기능이 변화된 폴리펩티드 변이체
US20020102208A1 (en) 1999-03-01 2002-08-01 Paul Chinn Radiolabeling kit and binding assay
CN1110322C (zh) 1999-07-21 2003-06-04 中国医学科学院医药生物技术研究所 单克隆抗体Fab'－平阳霉素偶联物及其抗肿瘤作用
WO2002002781A1 (en) 2000-06-30 2002-01-10 Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw Heterodimeric fusion proteins
GB0029407D0 (en) 2000-12-01 2001-01-17 Affitech As Product
ATE489395T1 (de) 2000-12-12 2010-12-15 Medimmune Llc Moleküle mit längeren halbwertszeiten, zusammensetzungen und deren verwendung
US7829084B2 (en) 2001-01-17 2010-11-09 Trubion Pharmaceuticals, Inc. Binding constructs and methods for use thereof
US20050107595A1 (en) 2001-06-20 2005-05-19 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
JP2005538706A (ja) 2001-07-12 2005-12-22 ジェファーソン フーテ， スーパーヒト化抗体
US6900292B2 (en) 2001-08-17 2005-05-31 Lee-Hwei K. Sun Fc fusion proteins of human erythropoietin with increased biological activities
US20040002587A1 (en) 2002-02-20 2004-01-01 Watkins Jeffry D. Fc region variants
US7317091B2 (en) 2002-03-01 2008-01-08 Xencor, Inc. Optimized Fc variants
AU2003209446B2 (en) 2002-03-01 2008-09-25 Immunomedics, Inc. Bispecific antibody point mutations for enhancing rate of clearance
GB0216648D0 (en) 2002-07-18 2002-08-28 Lonza Biologics Plc Method of expressing recombinant protein in CHO cells
DK1534335T4 (en) 2002-08-14 2015-10-05 Macrogenics Inc FCGAMMARIIB-SPECIFIC ANTIBODIES AND PROCEDURES FOR USE THEREOF
EP3502133A1 (en) 2002-09-27 2019-06-26 Xencor, Inc. Optimized fc variants and methods for their generation
AU2003286467B2 (en) 2002-10-15 2009-10-01 Abbvie Biotherapeutics Inc. Alteration of FcRn binding affinities or serum half-lives of antibodies by mutagenesis
EP2368578A1 (en) 2003-01-09 2011-09-28 Macrogenics, Inc. Identification and engineering of antibodies with variant Fc regions and methods of using same
JP4629047B2 (ja) 2003-06-12 2011-02-09 イーライ リリー アンド カンパニー Ｇｌｐ−１アナログ複合タンパク質
WO2005018572A2 (en) 2003-08-22 2005-03-03 Biogen Idec Ma Inc. Improved antibodies having altered effector function and methods for making the same
RU2392324C2 (ru) * 2003-09-18 2010-06-20 Симфоген А/С Способ связывания интересующих последовательностей
GB0324368D0 (en) 2003-10-17 2003-11-19 Univ Cambridge Tech Polypeptides including modified constant regions
BR122018071808B8 (pt) 2003-11-06 2020-06-30 Seattle Genetics Inc conjugado
CA2545603A1 (en) 2003-11-12 2005-05-26 Biogen Idec Ma Inc. Neonatal fc receptor (fcrn)-binding polypeptide variants, dimeric fc binding proteins and methods related thereto
US7001978B2 (en) 2003-11-19 2006-02-21 Xerox Corporation Unsaturated ester substituted polymers with reduced halogen content
US20050249723A1 (en) 2003-12-22 2005-11-10 Xencor, Inc. Fc polypeptides with novel Fc ligand binding sites
CN1918178B (zh) 2004-01-12 2012-08-22 应用分子进化公司 Fc区变体
EP1737890A2 (en) 2004-03-24 2007-01-03 Xencor, Inc. Immunoglobulin variants outside the fc region
WO2005123780A2 (en) 2004-04-09 2005-12-29 Protein Design Labs, Inc. Alteration of fcrn binding affinities or serum half-lives of antibodies by mutagenesis
WO2005111225A1 (en) 2004-05-04 2005-11-24 Novo Nordisk Health Care Ag O-linked glycoforms of polypeptides and method to manufacture them
WO2006085967A2 (en) 2004-07-09 2006-08-17 Xencor, Inc. OPTIMIZED ANTI-CD20 MONOCONAL ANTIBODIES HAVING Fc VARIANTS
EP2471813B1 (en) 2004-07-15 2014-12-31 Xencor, Inc. Optimized Fc variants
CA2580141C (en) 2004-09-23 2013-12-10 Genentech, Inc. Cysteine engineered antibodies and conjugates
AU2005289594B2 (en) 2004-09-27 2012-02-02 Centocor, Inc. Srage mimetibody, compositions, methods and uses
WO2006047350A2 (en) 2004-10-21 2006-05-04 Xencor, Inc. IgG IMMUNOGLOBULIN VARIANTS WITH OPTIMIZED EFFECTOR FUNCTION
JP4829609B2 (ja) * 2004-12-22 2011-12-07 独立行政法人科学技術振興機構 ヒト抗体酵素およびその生産方法
AU2006230413B8 (en) 2005-03-31 2011-01-20 Xencor, Inc Fc variants with optimized properties
JP2008537941A (ja) 2005-03-31 2008-10-02 ゼンコー・インコーポレイテッド 最適化特性を有するＦｃ変異体
WO2006130834A2 (en) 2005-05-31 2006-12-07 Board Of Regents, The University Of Texas System IGGl ANTIBODIES WITH MUTATED FC PORTION FOR INCREASED BINDING TO FCRN RECEPTOR AND USES THEREOF
US8901281B2 (en) 2005-06-17 2014-12-02 Merck Sharp & Dohme Corp. ILT3 binding molecules and uses therefor
SG163548A1 (en) 2005-06-30 2010-08-30 Centocor Inc Methods and compositions with enhanced therapeutic activity
US7612181B2 (en) 2005-08-19 2009-11-03 Abbott Laboratories Dual variable domain immunoglobulin and uses thereof
CZ297081B6 (cs) 2005-10-03 2006-09-13 Flek@Jaroslav Zubní kartácek
US7846724B2 (en) 2006-04-11 2010-12-07 Hoffmann-La Roche Inc. Method for selecting CHO cell for production of glycosylated antibodies
FI20060946A0 (fi) 2006-10-26 2006-10-26 Glykos Finland Oy Influenssaviruksen nukleiinihappoja ja peptidejä
JP2010512306A (ja) 2006-10-27 2010-04-22 ザ ロックフェラー ユニバーシティー 抗炎症特性が増強され、細胞毒性特性が減少したポリペプチドおよび関連する方法
CL2007003411A1 (es) 2006-11-28 2008-07-04 Centelion Proteina fusion que consiste en una region fc de una inmunoglobulina con un fragmento o dominio soluble de un receptor para fgf; polinucleotido que la codifica y vector y celula que lo comprenden; composicion farmaceutica que comprende la proteina fu
JP5357778B2 (ja) 2007-01-23 2013-12-04 ゼンコー・インコーポレイテッド 最適化ｃｄ４０抗体および前記を使用する方法
PE20090245A1 (es) 2007-05-08 2009-03-17 Genentech Inc Anticuerpos anti-muc16 disenados con cisteina y conjugados de anticuerpos y farmacos
US20100260751A1 (en) 2007-09-28 2010-10-14 Raju T Shantha Methods and Structural Conformations of Antibody Preparations with Increased Resistance to Proteases
EP2050764A1 (en) 2007-10-15 2009-04-22 sanofi-aventis Novel polyvalent bispecific antibody format and uses thereof
KR101622412B1 (ko) 2007-10-19 2016-05-18 제넨테크, 인크. 시스테인 조작된 항-ｔｅｎｂ２ 항체 및 항체 약물 접합체
CA2711503A1 (en) 2008-01-08 2009-07-16 Biogenerix Ag Glycoconjugation of polypeptides using oligosaccharyltransferases
CL2009000062A1 (es) 2008-01-31 2010-05-14 Genentech Inc Anticuerpo humanizado anti cd79b; con modificaciones de cisterna libre; inmunoconjugado que contiene dicho anticuerpo y una droga; polinucleotido que codifica el anticuerpo; vector, celula huesped; composicion farmaceutica y uso de dicha composicion para tratar cancer, preferentemente linfomas.
US20110137011A1 (en) 2008-04-21 2011-06-09 Novo Nordisk A/S Hyperglycosylated human coagulation factor ix
UA40611U (ru) 2008-05-26 2009-04-27 Владимир Семенович Живченко Способ десульфурации железоуглеродистого расплава в тигельной печи
EP2233499A1 (en) 2009-03-26 2010-09-29 CSL Behring AG Antibody composition with altered Fab sialylation
US10087236B2 (en) 2009-12-02 2018-10-02 Academia Sinica Methods for modifying human antibodies by glycan engineering
DK2603528T3 (en) 2010-08-10 2016-11-28 Glycotope Gmbh FAB-glycosylated ANTIBODIES
AR085302A1 (es) 2011-02-24 2013-09-18 Sanofi Sa Metodo de produccion de anticuerpos sialilados
SG193379A1 (en) 2011-03-11 2013-10-30 Amicus Therapeutics Inc Dosing regimens for the treatment of fabry disease
TWI838039B (zh) 2011-03-28 2024-04-01 法商賽諾菲公司 具有交叉結合區定向之雙重可變區類抗體結合蛋白
UY34317A (es) 2011-09-12 2013-02-28 Genzyme Corp Anticuerpo antireceptor de célula T (alfa)/ß
PT2895513T (pt) 2012-09-12 2018-10-08 Genzyme Corp Polipéptidos contendo fc com glicosilação alterada e função efetora reduzida
US9790268B2 (en) 2012-09-12 2017-10-17 Genzyme Corporation Fc containing polypeptides with altered glycosylation and reduced effector function
SG11201506088RA (en) 2013-03-11 2015-09-29 Genzyme Corp Hyperglycosylated binding polypeptides
HUE061339T2 (hu) 2014-03-19 2023-06-28 Genzyme Corp Targetáló csoportok helyspecifikus glikotechnikai átalakítása
EP3129402A4 (en) 2014-04-08 2017-12-13 University Of Georgia Research Foundation, Inc. Site-specific antibody-drug glycoconjugates and methods
AU2015330869B2 (en) 2014-10-09 2021-07-08 Genzyme Corporation Glycoengineered antibody drug conjugates
US10280317B2 (en) 2016-07-07 2019-05-07 Northwestern University Enhanced conductivity, adhesion and environmental stability of printed graphene inks with nitrocellulose

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0403156A1 (en) 1989-06-07 1990-12-19 Genzyme Corporation Improved monoclonal antibodies against the human alpha/beta t-cell receptor, their production and use
WO2004099249A2 (en) 2003-05-02 2004-11-18 Xencor, Inc. Optimized fc variants and methods for their generation
US20100190247A1 (en) 2003-12-04 2010-07-29 Xencor, Inc. Methods of Generating Variant Proteins with Increased Host String Content
WO2006105021A2 (en) 2005-03-25 2006-10-05 Tolerrx, Inc. Gitr binding molecules and uses therefor
US20080038260A1 (en) 2006-06-19 2008-02-14 Tolerrx, Inc. ILT3 binding molecules and uses therefor
WO2008086006A2 (en) 2007-01-09 2008-07-17 Biogen Idec Ma Inc. Sp35 antibodies and uses thereof
WO2010027797A1 (en) 2008-08-26 2010-03-11 Macrogenics Inc. T-cell receptor antibodies and methods of use thereof
WO2011109400A2 (en) 2010-03-04 2011-09-09 Macrogenics,Inc. Antibodies reactive with b7-h3, immunologically active fragments thereof and uses thereof

Non-Patent Citations (35)

* Cited by examiner, † Cited by third party
Title
Al-Lazikani et al. (J. Mol. Biol. (1997) 273, 927-948). *
Andrew C.R. Martin, "Protein Sequence and Structure Analysis of Antibody Variable Domains" in Chapter 3 of Antibody Engineering vol. 2, Kontermann and Dubel Eds., Springer-Verlag Berlin Heidelberg 2010, pp. 33-51. *
Chan et al. (2010) "Therapeutic antibodies for autoimmunity and inflammation," The Journal of Immunology. 10:301-316.
Feige et al., J. Mol. Biol. (2009) 391, 599-608. *
Giudicelli et al. (Cold Spring Harb Protoc. Jun. 1, 2001;2011(6), pp. 58-78). *
Heidecke et al. ‘Alpha-beta T Cell Receptor-directed Therapy in Rat Allograft Recipients’. 1996 Transplantation vol. 61 No. 6 pp. 948-956.
Heidecke et al. ‘Induction of Long-term Rat Renal Allograft Survival by Pretransplant T Cell Receptor Alpha-beta Targeted Therapy’. 1996 Transplantation vol. 61 No. 2 pp. 336-339.
Heidecke et al. 'Alpha-beta T Cell Receptor-directed Therapy in Rat Allograft Recipients'. 1996 Transplantation vol. 61 No. 6 pp. 948-956.
Heidecke et al. 'Induction of Long-term Rat Renal Allograft Survival by Pretransplant T Cell Receptor Alpha-beta Targeted Therapy'. 1996 Transplantation vol. 61 No. 2 pp. 336-339.
International Search Report. PCT/EP2012/003819 dated Jun. 28, 2013.
Jung et al. ‘Prevention and Therapy of Experimental Autoimmune Neuritis by an Antibody Against T Cell Receptors-alpha-beta’. 1992 Journal of Immunology vol. 148 No. 12 pp. 3768-3775.
Jung et al. 'Prevention and Therapy of Experimental Autoimmune Neuritis by an Antibody Against T Cell Receptors-alpha-beta'. 1992 Journal of Immunology vol. 148 No. 12 pp. 3768-3775.
Krapp et al., J. Mol. Biol. (2003) 325, 979-989. *
Labrijn et al. (2008) "When binding is enough: nonactivating antibody formats," Curr. Opin. Immunol. 20:479-485.
North et al. (J. Mol. Biol. (2011) 406, 228-256). *
Page et al. ‘Biologics in Organ Transplantation’. 2012 Transplant International. European Society for Organ Transplantation. vol. 25 pp. 707-719.
Page et al. 'Biologics in Organ Transplantation'. 2012 Transplant International. European Society for Organ Transplantation. vol. 25 pp. 707-719.
Sazinsky et al. (Proc Natl Acad Sci U S A. Dec. 23, 2008;105(51):20167-72 and Supplemental pp. 1-8). *
Sazinsky et al., PNAS, 2008, 105:51 20167-20172. *
Scharpf et al. ‘Immunomodulation with Anti-alpha-beta T Cell Receptor Monoclonal Antibodies in Combination With Cyclosporine A Improves Regeneration in Nerve Allografts’. 2006 Microsurgery vol. 26 pp. 599-607.
Scharpf et al. 'Immunomodulation with Anti-alpha-beta T Cell Receptor Monoclonal Antibodies in Combination With Cyclosporine A Improves Regeneration in Nerve Allografts'. 2006 Microsurgery vol. 26 pp. 599-607.
Schorlemmer et al. ‘Synergistic Effects of 15-Deoxyspergualin With Cyclosporine and the TCR-Targeted Monoclonal Antibody R73 to Induce Specific Unresponsiveness to Skin Allografts in Rats’ 1995 Transplantation Proceedings vol. 27 No. 1 pp. 414-416.
Schorlemmer et al. 'Synergistic Effects of 15-Deoxyspergualin With Cyclosporine and the TCR-Targeted Monoclonal Antibody R73 to Induce Specific Unresponsiveness to Skin Allografts in Rats' 1995 Transplantation Proceedings vol. 27 No. 1 pp. 414-416.
Shearman et al. ‘Construction, Expression, and Characterization of Humanized Antibodies Directed Against the Human Alpha-beta T Cell Receptor’. 1991 Journal of Immunology vol. 147 No. 12 pp. 4366-4373.
Shearman et al. 'Construction, Expression, and Characterization of Humanized Antibodies Directed Against the Human Alpha-beta T Cell Receptor'. 1991 Journal of Immunology vol. 147 No. 12 pp. 4366-4373.
Shields et al. (2001) "High Resolution Mapping of the Binding Site on Human IgG1 for FcyRl, FcyRll, FcyRlll, and FcRn and Design of IgG1 Variants with Improved Binding to the FcyR," J. Biol. Chem. 276:6591-6604.
Williams et al. ("Humanising Antibodies by CDR Grafting" in Chapter 21 of Antibody Engineering vol. 1, Kontermann and Dubel Eds., Springer-Verlag Berlin Heidelberg 2010, pp. 319-339). *
Wright et al. ‘Genetically Engineered Antibodies: Progress and Prospects’. 1992 Critical Reviews in Immunology vol. 12 No. 3-4 pp. 125-168.
Wright et al. 'Genetically Engineered Antibodies: Progress and Prospects'. 1992 Critical Reviews in Immunology vol. 12 No. 3-4 pp. 125-168.
Written Opinion of The International Searching Authority. PCT/EP2012/003819 dated Sep. 12, 2011.
Yamagami et al. ‘Suppression of Allograft Rejection with Anti-alpha-beta T Cell Receptor Antibody in Rat Corneal Transplantation’. 1999 Transplantation vol. 67 No. 4 pp. 600-604.
Yamagami et al. 'Suppression of Allograft Rejection with Anti-alpha-beta T Cell Receptor Antibody in Rat Corneal Transplantation'. 1999 Transplantation vol. 67 No. 4 pp. 600-604.
Yoshino, Shin. Cleland, Leslie G. ‘Depletion of Alpha-beta T Cells by a Monoclonal Antibody against the Alpha-beta T Cell Receptor Suppresses Established Adjuvant Arthritis, but not Established Collagen-induced Arthritis in Rats’. 1992 J. Exp. Med. vol. 175 pp. 907-915.
Yoshino, Shin. Cleland, Leslie G. 'Depletion of Alpha-beta T Cells by a Monoclonal Antibody against the Alpha-beta T Cell Receptor Suppresses Established Adjuvant Arthritis, but not Established Collagen-induced Arthritis in Rats'. 1992 J. Exp. Med. vol. 175 pp. 907-915.
Zhou et al. (2008) "Development of a simple and rapid method for producing non-fucosylated oligomannose containing antibodies with increased effector function," Biotechnol. Bioeng. 99:652-665.

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* Cited by examiner, † Cited by third party
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US11807690B2 (en) 2013-03-11 2023-11-07 Genzyme Corporation Hyperglycosylated binding polypeptides
US12110338B2 (en) 2013-03-11 2024-10-08 Genzyme Corporation Site-specific antibody-drug conjugation through glycoengineering
US20220098299A1 (en) * 2013-10-30 2022-03-31 Genzyme Corporation Methods for enhancing immunosuppressive therapy by multiple administration of alpha beta tcr-binding polypeptide
US11697690B2 (en) 2014-03-19 2023-07-11 Genzyme Corporation Site-specific glycoengineering of targeting moieties

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